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Plant surfaces are the interfaces of the organisms with respect to their environment. In the micro-dimension they show an enormous variety of functional three-dimensional structures. Their materials and structures developed over millions of years by evolutionary processes in which their functionality has been proven and selected by environmental pressures. As a result, nature developed highly functional materials with several amazing properties like superhydrophobicity and superhydrophilicity. These functional structures are built up by a complex biopolymer called cuticle. The cuticle is mainly composed of a three-dimensional network of cutin, and integrated and superimposed lipids called "waxes". Superimposed waxes are also called "epicuticular waxes". Epicuticular waxes often form two- and three-dimensional structures, in dimensions between hundreds of nanometers and some micrometers, which influence the wettability, self-cleaning behaviour and the light reflection at the cuticle interface. This review gives a brief introduction into the functions of the plant epicuticular waxes and summarises the current knowledge about their morphologies, crystal structures, growth by self-assembly and provides an overview about the microscopy and preparation techniques for their analysis.

The crystal structure of the epicuticular waxes of 35 plant species has been examined by electron diffraction and X-ray powder diffraction. The waxes include the most common morphological wax types such as platelets, tubules, films and rodlets. Most of them were prepared with a special mechanical isolation method, which preserves the original crystal structure. Solvent-extracted recrystallized plant waxes were compared with mechanically isolated samples. The waxes were found to occur in three different crystal structures. Most of the waxes exhibited an orthorhombic structure which is the most common for aliphatic compounds. Tubules containing mainly secondary alcohols showed diffraction reflections of a triclinic phase; broad reflection peaks indicated a significant disorder. Ketones, in particular beta-diketone tubules, displayed the reflections of a hexagonal structure. Mixtures of different phases could be identified. For most of the waxes, the 'long spacing' diffraction reflections indicated a layer structure with the characteristics of the major component. Others showed no 'long spacing' reflections indicating a strong disorder of the molecular layers.

The cuticle of terrestrial vascular plants and some bryophytes is covered with a complex mixture of lipids, usually called epicuticular waxes. Self-assembly processes of wax molecules lead to crystalline three-dimensional micro- and nanostructures that emerge from an underlying wax film. This paper presents the first AFM study on wax regeneration on the surfaces of living plants and the very early stages of wax crystal formation at the molecular level. Wax formation was analysed on the leaves of Euphorbia lathyris, Galanthus nivalis, and Ipheion uniflorum. Immediately after wax removal, regeneration of a wax film began, consisting of individual layers of, typically, 3-5 nm thickness. Subsequently, several different stages of crystal growth could be distinguished, and different patterns of wax regeneration as well as considerable variation in regeneration speed were found.